人类ARRB1和GNMT蛋白相互作用的初步研究  

Preliminary Research on the Interaction between GNMT and ARRB1

作　　者：叶侃[1] 石彦[1] 伍慧玲[1] 周娟[1] 施慧莉[1] 霍克克[1] 

机构地区：[1]复旦大学生命科学学院遗传学研究所遗传工程国家重点实验室,上海200433

出　　处：《复旦学报（自然科学版）》2010年第1期29-35,42,F0003,共9页Journal of Fudan University：Natural Science

基　　金：国家高技术研究发展计划(863计划)资助项目(2006AA02A310);上海市科技创新团队计划资助项目(03DZ14024)

摘　　要：GNMT(甘氨酸-N-甲基转移酶,glycine N-methyltransferase)是人体内一个多功能蛋白.最近研究发现GNMT是一个潜在的抑癌基因,但GNMT抑癌的机理却不清楚.研究以GNMT为"诱饵",利用酵母双杂交系统筛选人肝cDNA文库,获得了GNMT的一个相互作用蛋白ARRB1(-βarrestin1),并通过GST Pull-down、免疫共沉淀的方法在体内体外验证了相互作用的特异性.进一步研究发现GNMT和ARRB1的相互作用不依赖于GNMT的四聚体高级结构和甲基转移酶活性.免疫荧光共定位实验发现,当ARRB1与GNMT共转染HeLa细胞后,ARRB1蛋白的亚细胞定位发生改变,表现出与GNMT蛋白共定位,有入核的趋势.这为研究GNMT的抑癌作用机理提供了新的线索.GNMT （glycine N-methyltransferase） is a multifunctional protein. Recent studies showed that GNMT may be a potential tumor suppressor gene. However, the mechanism is unclear yet. Here, GNMT was used as a ＂bait＂ to screen a human liver eDNA library and ARRB1 was identified as a GNMT binding protein. Their special interaction was confirmed by GST pull-down and co-immunoprecipitation assays. Further, the interaction was found to be independent of the tetrameric structure and enzyme activity of GNMT. Immunofluorescent localization experiments showed that ARRB1 translocated from cytoplasm to nucleus when co-transformed with GNMT in HeLa cells so as to co-localize with GNMT. These results provided a clue to explore the role of GNMT in tumorigenesis.

关 键 词：GNMT ARRB1 酵母双杂交 免疫荧光定位 

分 类 号：Q71[生物学—分子生物学]